Rapid and sensitive screening and confirmation of thirty‐four aminocarbonyl/carboxamide (NACA) and arylindole synthetic cannabinoid drugs in human whole blood

An autopsy case in which the cause of death was judged as drug poisoning by two synthetic cannabinoids, including MAB-CHMINACA, was investigated. Although unchanged MAB-CHMINACA could be detected from solid tissues, blood and stomach contents in the case, the compound could not be detected from a urine specimen. We obtained six kinds of reference standards of MAB-CHMINACA metabolites from a commercial source. The MAB-CHMINACA metabolites from the urine specimen of the abuser were extracted using a QuEChERS method including dispersive solid-phase extraction, and analyzed by liquid chromatography-tandem mass spectrometry with or without hydrolysis with β-glucuronidase. Among the six MAB-CHMINACA metabolites tested, two predominant metabolites could be identified and quantified in the urine specimen of the deceased. After hydrolysis with β-glucuronidase, an increase of the two metabolites was not observed. The metabolites detected were a 4-monohydroxycyclohexylmethyl metabolite M1 (N-(1-amino-3,3-dimethyl-1-oxobutan-2-yl)-1-((4-hydroxycyclohexyl)methyl)-1H-indazole-3-carboxamide) and a dihydroxyl (4-hydroxycyclohexylmethyl and tert-butylhydroxyl) metabolite M11 (N-(1-amino-4-hydroxy-3,3-dimethyl-1-oxobutan-2-yl)-1-((4-hydroxycyclohexyl)methyl)-1H-indazole-3-carboxamide). Their concentrations were 2.17 ± 0.15 and 10.2 ± 0.3 ng/mL (n = 3, each) for M1 and M11, respectively. Although there is one previous in vitro study showing the estimation of metabolism of MAB-CHMINACA using human hepatocytes, this is the first report dealing with in vivo identification and quantification of MAB-CHMINACA metabolites in an authentic human urine specimen.

Section: Introductionmentioning

confidence: 99%

Identification and quantification of predominant metabolites of synthetic cannabinoid MAB‐CHMINACA in an authentic human urine specimen

Hasegawa

Minakata

Gonmori

et al. 2017

“…22,23 MDMB-CHMINACA has also been included in the testing scope of various laboratories in serum and whole blood. 24,25 Two other synthetic cannabinoids, MDMB-CHMICA and MAB-CHMINACA (ADB-CHMINACA) ( Figure 1), are structurally related to MDMB-CHMINACA and have been identified internationally and reported in peer-reviewed literature. MDMB-CHMICA is an indole carboxamide, differing from MDMB-CHMINACA only in that the core ring is indole instead of indazole.…”

Section: Mdmb-chminacamentioning

confidence: 99%

“…Additional studies have been conducted on MDMB‐CHMINACA, including its mass spectral characteristics in electrospray and electron ionization sources, as well as its stability in serum extracts under various laboratory storage conditions . MDMB‐CHMINACA has also been included in the testing scope of various laboratories in serum and whole blood . Two other synthetic cannabinoids, MDMB‐CHMICA and MAB‐CHMINACA (ADB‐CHMINACA) (Figure ), are structurally related to MDMB‐CHMINACA and have been identified internationally and reported in peer‐reviewed literature.…”

Section: Introductionmentioning

confidence: 99%

In vitro Phase I metabolism of indazole carboxamide synthetic cannabinoid MDMB‐CHMINACA via human liver microsome incubation and high‐resolution mass spectrometry

Presley

Logan

Jansen‐Varnum

2019

Self Cite

Synthetic cannabinoids have proliferated over the last decade and have become a major public health and analytical challenge, critically impacting the clinical and forensic communities. Indazole carboxamide class synthetic cannabinoids have been particularly rampant, and exhibit severe toxic effects upon consumption due to their high binding affinity and potency at the cannabinoid receptors (CB1 and CB2). MDMB‐CHMINACA, methyl 2‐[1‐(cyclohexylmethyl)‐1H‐indazole‐3‐carboxamido]‐3,3‐dimethylbutanoate, a compound of this chemical class, has been identified in forensic casework and is structurally related to several other synthetic cannabinoids. This study presents the first extensive report on the Phase I metabolic profile of MDMB‐CHMINACA, a potent synthetic cannabinoid. The in vitro metabolism of MDMB‐CHMINACA was determined via incubation with human liver microsomes and high‐resolution mass spectrometry. The accurate masses of precursor and fragments, mass error (ppm), and chemical formula were obtained for each metabolite. Twenty‐seven metabolites were identified, encompassing twelve metabolite types. The major biotransformations observed were hydroxylation and ester hydrolysis. Hydroxylations were located predominantly on the cyclohexylmethyl (CHM) moiety. Ester hydrolysis was followed by additional biotransformations, including dehydrogenation; mono‐ and dihydroxylation and ketone formation, each with dehydrogenation. Minor metabolites were identified and reported. The authors propose that CHM‐monohydroxylated metabolites specific to MDMB‐CHMINACA are the most suitable candidates for implementation into bioanalytical assays to demonstrate consumption of this synthetic cannabinoid. Due to the structural similarity of MDMB‐CHMINACA and currently trending synthetic cannabinoids whose metabolic profiles have not been reported, the results of this study can be used as a guide to predict their metabolic pathways.

“…A lot of specific methods for the determination of synthetic cannabinoids in whole blood or serum were described. Kacinko et al .…”

Section: Introductionmentioning

confidence: 99%

“…A lot of specific methods for the determination of synthetic cannabinoids in whole blood [1][2][3] or serum [4][5][6][7] were described. Kacinko et al [1] showed that JWH-018, JWH-073, and JWH-250 were stable at room, refrigerated, and frozen temperatures for at least 30 days in whole blood containing EDTA.…”

Section: Introductionmentioning

confidence: 99%

Freeze‐thaw stability and long‐term stability of 84 synthetic cannabinoids in serum

Heß

Krueger

Unger

et al. 2016

Information about stability of synthetic cannabinoids is important to give recommendations for storage conditions in cases in which use of synthetic cannabinoids is suspected. In this study, freeze-thaw stability (3 cycles at -20 °C at 1.5 ng/mL) and long-term stability at room temperature, 4°C and -20°C (for 150 days) were tested by a validated liquid chromatographic-mass spectrometric method for 80 synthetic cannabinoids. Results demonstrated good freeze-thaw stability for most of the substances. For 5 F-ABICA (72.6% of peak area of time point zero), 5 F- 2 ADB PINACA 2 Isomer (68.7 %), 5Cl-AB-PINACA (84.2 %), AB FUBINACA 2-Isomer (33.5 %), 5 F-PCN (89.7 %), ADB-FUBINACA (78.0 %), EG 018 (88.9 %), and PX-1 (89.4 %) three freeze-thaw cycles led to absolute peak areas < 90% compared to 6 control samples. For 51 of the 84 substances, stability in serum could be demonstrated after 315 days of storage at -20°C. For 82 of the 84 substances, stability in serum could be shown for at least 1 month at -20°C. Long- term stability at 4°C or 20°C showed worse results. Therefore, it is our recommendation to store real serum samples at -20°C until analysis. Copyright © 2016 John Wiley & Sons, Ltd.